ABSTRACT: In the past decade several alternative vaping products have hit the market, rapidly gaining consumers among adults and, especially, adolescents. Electronic nicotine delivery systems or e-cigarettes (e-Cigs) have become wide spread among both smokers and former non-smokers due to the belief that they are much safer than traditional cigarettes. Among them JUULs have become the sought-after product among youth including middle schoolers. Moreover, it is well established that tobacco smoking (TS) has been associated with vascular endothelial dysfunction in a causative and dose dependent manner primarily related to the TS content of reactive oxygen species (ROS), nicotine, and oxidative stress (OS) -driven inflammation. Current scientific opinion considers OS-mediated pathways to play a major role in the pathogenesis of these disorders, especially stroke. Preclinical studies have also shown that nicotine (the principal e- liquid's ingredient used in e-cigarettes) can also cause OS, exacerbation of cerebral ischemia and secondary brain injury. Likewise, chronic e-Cig vaping could be prodromal to cerebrovascular impairment and promote cerebrovascular conditions favoring the onset of stroke and worsening post-ischemic brain injury. The health impact associated with chronic e-Cig vaping is largely unknown especially concerning the cerebrovascular system. Wide spread use of these alternative products strongly calls for investigative studies to determine their real health impact as well as more stringent regulatory guidelines concerning the content of the vaping solutions (e-liquids). Our new preliminary data also suggests the possibility for gender specific sensitivities to these products. Thus, in response to the program scope and research objectives of this call for action to investigate e-Cig health effects and toxicity issued by the FDA Center for Tobacco Products Based and based on the substantial published and preliminary data from our Labs we propose the following: 1) Assess the cerebrovascular impact of e-Cig vaping and Juuling vs. TS exposure and develop a panel of potential biomarkers to determine harm of these products. The premise of these studies is to comparatively investigate side by side the harm or toxicity of e-Cigs and JUUL vs. TS on the BBB. This includes assessing the impact of different e-liquid formulations in respect to the nicotine content as well as impact of the coil heat used to generate the vapor. 2) Evaluate and validate in vivo the impact of chronic exposure to e-Cigs and JUULs vs. TS on the risk of stroke, secondary brain damage, and post-ischemic neurological impairments. The premise for these studies builds upon the latest clinical findings and our recent published data suggesting that similarly to TS, chronic e-Cigs exposure promotes cerebrovascular inflammation, stroke, and worsen post-ischemic secondary brain injuries. To also address the possibility for gender specific sensitivities/risks and outcomes we will use a mixed gender population of male and female mice. Overall, we will compare the impact of tobacco smoking and e-Cig vaping (including e-liquids containing different nicotine concentrations and temperature settings used to vaporize the e-liquid) on brain vascular damage. Our study will focus specifically on their impact on stroke risk and outcome, ascertained from brain and blood based biomarkers.